Katherine Excoffon (Committee Member), Chandler Phillips (Committee Member), Jaime Ramirez-Vick (Advisor)
Master of Science in Biomedical Engineering (MSBME)
The use of the standard two dimensional (2D) cell culture has laid down the fundamentals of molecular and cell biology. However, recent advances in cell-based regenerative medicine raises the concern on deconstructing cellular behaviors in more physiologically relevant three dimensional (3D) microenvironments. Differences in cell response in 2D versus 3D systems arise from the perturbations in gene expression patterns that stem from how cells sense their underlying 2D or surrounding 3D matrices and adjust their phenotypes accordingly. Thus, cells are no longer considered as a solitary entity of genome but a context arises from a combinatorial interactions of cell-ECM, cell-cell, and cell-biomolecules that constitutively organize the tissues and ultimate functional organs. In this regard, this project was intended to identify cell behaviors and gene expression pattern in 2D versus 3D culture systems. To construct 2D versus 3D cultures of human adipose tissue-derived mesenchymal stem cells (hASCs) were seeded on the top of plastic substrates or encapsulated in the self-assembled polypeptide-based hydrogel PuraMatrix™, respectively. Adipogenic, osteogenic, and chondrogenic differentiation in these two different culture systems was induced with the corresponding lineage-specific biochemical induction medium. The effect on growth and differentiation of hASCs under these two culture conditions was assessed through changes in cellular morphology, survivability, proliferation, and gene expression pattern during lineage-specific differentiation. According to the cytotoxicity assay, PuraMatrix™ provides a suitable 3D microenvironment for hASCs proliferation and differentiation without the need of any exogeneous cell adhesion motifs, however an influence of initial cell seeding density was seen in maintaining high initial cell viability and subsequent functionality. Gene microarray analysis revealed that, compared to 2D culture, 3D culture shows more of the expected lineage-specific gene expression and supports tissue specific developmental characteristics.
Department or Program
Department of Biomedical, Industrial & Human Factors Engineering
Year Degree Awarded
Copyright 2018, all rights reserved. My ETD will be available under the "Fair Use" terms of copyright law.